KR20010012812A - Compounds for immunotheraphy and diagnosis of tuberculosis and methods of their use - Google Patents

Abstract

The present invention relates to compositions and methods for inducing prophylactic immunity against tuberculosis. Compositions of the present invention comprise a polypeptide comprising at least one immunogenic portion of at least one M. tuberculosis protein and a DNA molecule encoding the polypeptide. Such compositions may be formulated with vaccines and / or pharmaceutical compositions for immunization against M. tuberculosis infection and may also be used to identify tuberculosis.

Description

COMPOUNDS FOR IMMUNOTHERAPHY AND DIAGNOSIS OF TUBERCULOSIS AND METHODS OF THEIR USE}

Tuberculosis is a chronic infectious disease usually caused by infection with Mycobacterium tuberculosis. Tuberculosis is not only a major disease in developing countries but also an increasing problem in developed countries around the world, with about 8 million cases occurring, killing 3 million people every year. Although infection does not cause symptoms for a significant period of time, most commonly the disease manifests as acute inflammation of the lung resulting in fever and an unproductive cough. If left untreated, it can lead to serious complications and usually death.

Tuberculosis can generally be controlled by extending the use of antibiotic therapy, but such treatment is not sufficient to prevent the spread of the disease. Infected individuals may be asymptomatic, but are sometimes contagious. In addition, compliance with the therapy is important, but it is difficult to monitor patient behavior. Some patients may cause ineffective treatment and tolerability without completing the course of treatment.

To suppress the spread of tuberculosis requires effective vaccination and accurate early identification of the disease. Currently, vaccination with live bacteria is the most effective way to induce protective immunity. The most common mycobacterium used for this purpose is Baccillus Calmette-Guerin (BCG), a non-toxic strain of Mycobacterium bovis. However, the safety and efficacy of BCG are controversial and some countries, such as the United States, do not receive the vaccine for the general public. Identification is usually done using a skin test that is exposed intradermal to tuberculin PPD (protein purified derivatives). An antigen specific T cell response results in measurable cirrhosis at the injection site 48-72 hours after injection, indicating that the cirrhosis has been exposed to the antigen of mycobacteria. However, problems with this test method are sensitivity and specificity, and individuals inoculated with BCG are indistinguishable from infected individuals.

Macrophages are M. It appears to act as a basic effector of tuberculous immunity and T cells are the major inducers of this immunity. M. The essential role of T cells in the protection against tuberculous infections is that M. tuberculosis occurs frequently in AIDS patients due to the depletion of CD4 T cells associated with human immunodeficiency virus (HIV) infection. It can be seen that. Mycobacterium reactive CD4 T cells appear to be effective producers of gamma interferon (IFN-γ), and IFN-γ also appears to trigger the mycobacterial inhibitory effect of macrophages in mice. The role of IFN-γ in humans is less clear than in mice, but from several studies 1,25-dihydroxy-vitamin D3, alone or in combination with IFN-γ or tumor necrosis factor-alpha, has been shown to inhibit human macrophages. Activation was shown to inhibit M. tuberculosis infection. In addition, IFN- [gamma] is known to stimulate human macrophages to produce 1,25-dihydroxy-vitamin D3. Similarly, IL-12 has been shown to play a role in stimulating resistance to M. tuberculosis infection. For immunology of M. tuberculosis infection, see Chan and Kaufmann et al., Tuberculosis: Pathogenesis, Protection and Control, Bloom (ed.), ASM Press, Washington, DC, 1994.

Thus, there is a need in the art for improved vaccines and methods for preventing, treating and detecting tuberculosis. The present invention satisfies this need and further provides other related advantages.

The present invention generally relates to methods for detecting, treating and preventing Mycobacterium tuberculosis infections. More specifically, the invention provides a polypeptide comprising a mycobacterium tuberculosis antigen, or part or other variant thereof, and a polypeptide for use in the identification and vaccination of mycobacterium tuberculosis infection. It is about.

1A and 1B show stimulation of proliferation and interferon-γ production in T cells derived from a first PPD positive donor (hereinafter referred to as D7) by synthetic peptides for recombinant ORF-2 and ORF-2, respectively.

2A and 2B show stimulation of proliferation and interferon-γ production in T cells derived from a second PPD positive donor (hereinafter referred to as D160) by synthetic peptides for recombinant ORF-2 and ORF-2, respectively.

Detailed description of the invention

As noted above, the present invention generally relates to compositions and methods for preventing, treating and identifying tuberculosis. The composition of the present invention is M. A polypeptide comprising a variant of said antigen that has been altered by one or more immunogenic portions of the tuberculosis antigen or only by conservative substitutions and / or modifications. The term "polypeptide" herein is a concept comprising an amino acid chain of any length, including a full-length protein (ie, an antigen) to which amino acid residues are joined by peptide covalent bonds. Thus, a polypeptide comprising an immunogenic portion of one of the antigens may consist only of the immunogenic portion or may comprise additional sequences. Additional sequences are native M. a. It may be derived from a tuberculosis antigen or may be heterologous and this sequence may be immunogenic (but not required).

"Immunogenic" as used herein refers to the ability to induce an immune response (eg, a cellular immune response) in a patient, such as a human, and / or in a biological sample. Specifically, antigens that are immunogenic (and immunogenic portions or other variants of these antigens) are one or more cells selected from the group consisting of T cells, NK cells, B cells, and macrophages (these cells are M. tuberculosis immunity). Cell proliferation, interleukin-12 production, and / or interferon- [gamma] production in biological samples, including from a subject). At least one M. Polypeptides comprising one or more immunogenic portions of the tuberculosis antigen can generally be used to detect tuberculosis in patients or to induce prophylactic immunity against tuberculosis.

The compositions and methods of the present invention also include variants of the polypeptides. As used herein, a polypeptide “variant” is a polypeptide that differs from the polypeptide mentioned only in conservative substitutions and / or modifications and which possesses the therapeutic, antigenic and / or immunogenicity of the polypeptide. Polypeptide variants exhibit at least about 70%, more preferably at least about 90%, most preferably at least 95% identity with the identified polypeptides. For polypeptides with immune reactivity, the variant may also be identified by modifying the amino acid sequence of one of the polypeptides and evaluating the immune reactivity of the modified polypeptide. In the case of polypeptides useful for the production of identifying binders, the variant can be identified by assessing the ability of the modified polypeptide to produce antibodies that detect the presence or absence of tuberculosis. On the other hand, variants of the antigens claimed herein that can be usefully used in the identification method of the present invention can be identified by evaluating the ability of the modified polypeptide to detect antibodies present in the serum of patients infected with tuberculosis. Such modified sequences can be prepared and tested using, for example, the representative procedures described herein.

"Conservative substitutions" refers to the substitution of an amino acid with another amino acid with similar properties, such that one skilled in the art of peptide chemistry can predict that the secondary structure and hydrotherapeutic properties of the polypeptide are substantially unchanged. In general, the following groups of amino acids show conservative changes: (1) ala, pro, gly, glu, asp, gln, asn, ser, thr; (2) cys, ser, tyr, thr; (3) val, ile, leu, met, ala, phe; (4) lys, arg, his; And (5) phe, tyr, trp, his.

Variants can also be modified by methods such as deletion or addition of amino acids with minimal impact on the immunogenicity, secondary structure and hydropathic of the polypeptide. For example, the polypeptide may be conjugated to the N-terminal signal (or leader) sequence of the protein which controls the delivery of the protein simultaneously with or after translation. The polypeptide may also be conjugated to a linker or other sequence for ease of synthesis, purification or identification of the polypeptide (eg, poly-His) or to enhance binding of the polypeptide to a solid support. For example, the polypeptide can be conjugated to an immunoglobulin Fc region.

In general, M. The tuberculosis antigen and the DNA sequence encoding the antigen can be prepared using any of a variety of methods. For example, M. The genome or cDNA library from tuberculosis may contain one or more M. a. Peripheral blood mononuclear cells (PBMCs) derived from tuberculosis immune individuals, T cell lines or T cell clones can be used for direct selection. Direct library screening is usually M. For the ability to induce proliferation and / or interferon- [gamma] production in T cells derived from a tuberculosis immune individual, a pool of expressed recombinant proteins can be performed. As described above, latent T cell antigens may be selected first based on antibody reactivity.

Meanwhile, the DNA sequence encoding the antigens is M. a. Suitable M. using serum from patients infected with tuberculosis. The tuberculous genome or cDNA expression library can be selected and identified. Such screening can generally be performed using techniques well known to those skilled in the art, such as those described in Sambrook et al., Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratories, Cold Spring Harbor, New York, 1989.

Purified antigens are assessed for their ability to elicit a suitable immune response (eg, cellular immune response) using, for example, the representative methods disclosed herein. The immunogenic antigen can then be partially sequenced using techniques such as traditional Edman chemistry. Edman and Berg, Eur. J. Biochem. 80: 116-132, 1967. Immunogenic antigens can also be prepared recombinantly using DNA sequences encoding antigens, wherein the DNA sequences are inserted into expression vectors and expressed in a suitable host.

DNA sequences encoding antigens of the present invention may also be suitable for DNA sequences that hybridize to degenerate oligonucleotides derived from partial amino acid sequences of isolated antigens. It can also be obtained by selection from tuberculosis cDNA or genomic DNA libraries. Degenerate oligonucleotide sequences used for such selection are designed as described in Sambrook et al., Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratories, Cold Spring Harbor, New York, 1989, and references cited therein, Can be synthesized and screening can be performed. Nucleic acid probes can also be isolated from cDNA or genomic libraries using polymerase chain reaction (PCR) using such oligonucleotides by methods well known in the art. Library selection can then be performed using isolated probes.

Regardless of the method of preparation, the antigens (and immunogenic portions thereof) disclosed herein have the ability to induce immunogenic responses. More specifically, the antigen is M. a. Has the ability to induce proliferation and / or cytokine production (ie, interferon-γ and / or interleukin-12 production) in T cells, NK cells, B cells and / or macrophages derived from a tuberculosis immune entity . The choice of cell type for use in assessing an immunogenic response to an antigen depends on the desired response. For example, interleukin-12 production is most readily assessed using agents containing B cells and / or macrophages. M. Tuberculous immunity object is M. Refers to an individual (substantially free of signs of disease) that is considered resistant to the development of tuberculosis by causing an effective T cell response to tuberculosis. Such individuals may be identified based on the strong intradermal skin test response to tuberculosis protein (PPD) (ie, indicating cirrhosis greater than about 10 mm in diameter) and the absence of any signs or signs of tuberculosis. M. T cells, NK cells, B cells and macrophages from tuberculosis-immune individuals can be prepared using methods known to those skilled in the art. For example, preparations of PBMCs (ie, peripheral blood mononuclear cells) can be used without further isolation of constituent cells. In general, PBMCs can be prepared using density centrifugation, such as through Ficoll ^™ (Winddrop Laboratories, NY).

In addition, T cells for use in the assays described herein can also be purified directly from PBMC. Alternatively, T cell clones reactive to mycobacterial proteins or T cell clones reactive to mycobacterial proteins of the individual may be used. Such T cell clones are described, for example, in M. a. PBMCs derived from tuberculosis immune individuals can be produced by incubating with mycobacterial proteins for 2-4 weeks. This process multiplies only mycobacterial protein-specific T cells to produce a cell line consisting only of those cells. Individual proteins can be cloned and tested using methods known to those of skill in the art to more accurately confirm the T cell specificity of the individual. In general, M. Tests in assays for proliferation and / or cytokine production (ie, interferon-γ and / or interleukin-12 production) using T cells, NK cells, B cells and / or macrophages derived from tuberculosis immune individuals Antigens carried out in this positive are considered to be immunogenic. This analysis can be performed using, for example, the representative procedure described below. Immunogenic portions of such antigens can be identified using similar assays and can be present in the polypeptides disclosed herein.

The ability of a polypeptide (eg, an immunogenic antigen or portion thereof or other variant) to induce cell proliferation is assessed by contacting the cells (eg, T cells and / or NK cells) with the polypeptide and measuring the proliferation of these cells. . Generally, the amount of polypeptide sufficient to assess about 10 ⁵ cells is about 10 ng / ml to about 100 μg / ml, preferably about 10 μg / ml. Incubation of cells and polypeptides is typically performed at 37 ° C. for about 6 days. After incubation with the polypeptide, cells are assayed for proliferative response, which method comprises methods known in the art, such as exposing the cells to radiolabeled thymidine pulses and measuring the label entrapped with intracellular DNA. It can be evaluated by a method including a step. In general, polypeptides that result in a threefold or more increase in background proliferation (ie, proliferation observed for cells cultured without polypeptide) are considered to induce proliferation.

The ability of a polypeptide to stimulate the production of interferon- [gamma] and / or interleukin-12 intracellularly comprises contacting the polypeptide with the cell and measuring the amount of interferon- [gamma] or interleukin-12 produced by the cell. Can be evaluated. Generally, the amount of polypeptide sufficient to assess about 10 ⁵ cells is about 10 ng / ml to about 100 μg / ml, preferably about 10 μg / ml. The polypeptide may be immobilized on a solid support such as beads or on biodegradable microspheres such as those disclosed in US Pat. Nos. 4,897,268 and 5,075,109, but need not be so. Incubation of cells and polypeptides can usually be performed at 37 ° C. for about 6 days. After incubation with the polypeptide, the cells are analyzed for interferon-γ and / or interleukin-12 (or one or more subunits), wherein methods known in the art, such as enzyme-linked immunosorbent (ELISA) or In the case of IL-12 P70 heterodimer, it can be evaluated by bioanalytical methods such as measuring the proliferation of T cells. In general, the culture supernatant (1 ㎖ per 10 ⁵ T cells containing ~10 ⁴⁾ 1 ㎖ per interferon polypeptide generating -γ 50 pg or more can be seen to stimulate the production of interferon -γ. Polypeptides that stimulate the production of at least 10 pg / ml of IL-12 P70 subunits and / or at least 100 pg / ml of IL-12 P40 subunits per 10 ⁵ macrophages or B cells (or per 3 × 10 ⁵ PBMCs) It can be seen to stimulate the production of -12.

In general, immunogenic antigens refer to proliferation and / or cytokine production (ie interferon-γ and / or in T cells, NK cells, B cells and / or macrophages derived from at least about 25% of M. tuberculosis immune individuals). Or interleukin-12 production). Among these immunogenic antigens, polypeptides having excellent therapeutic properties can be distinguished based on the extent of response and the proportion of individuals for whom a response was observed in the assay. In addition, antigens with good therapeutic properties do not stimulate in vitro proliferation and / or cytokine production in cells induced in at least about 25% of individuals without M. tuberculosis immunity, and therefore specifically M. tuberculosis Eliminate reactions that are not due to reactive cells. These antigens that induce responses at high rates of T cells, NK cells, B cells and / or macrophage preparations derived from M. tuberculosis immune individuals (individuals with a low incidence of responses in cell production from other individuals) Excellent therapeutic properties.

Antigens with good therapeutic properties can be identified based on their ability to reduce the intensity of M. tuberculosis infection in experimental animals when administered with a vaccine. Vaccine formulations suitable for experimental animals are described in detail below. Efficacy may be determined based on the ability of a constant temperature to provide at least about 50% reduction in bacterial count and / or at least about 40% reduction in mortality after experimental infection. Suitable experimental animals include mice, guinea pigs and primates.

In general, antigens with good identification properties can induce a response in an intradermal skin test performed in an individual with active tuberculosis but not in a test performed in an individual not infected with M. tuberculosis. You can identify yourself based on your abilities. Skin tests can generally be performed as described below, but are considered positive if they exhibit a cirrhosis response of 5 mm or greater.

The immunogenic portion of the antigens disclosed herein are prepared using known techniques, such as those outlined in Paul et al., Fundamental Immunology, 3d ed., Raven Press, 1993, p243-247, and the references cited herein. And identifiable. Such techniques include searching for polypeptide portions of natural antigens that have immunogenic properties. Representative proliferation and cytokine production assays disclosed herein can generally be used at screening. The immunogenic portion of a polypeptide is the portion that produces an immune response (eg, proliferation, interferon-γ production, and / or interleukin-12 production) that is nearly similar to that of a full-length antigen in a typical assay. That is, the immunogenic portion of the antigen can produce at least about 20%, preferably about 100%, of the full-length antigen-induced proliferation in the model proliferation assays disclosed herein. The immunogenic portion can also stimulate the production of at least about 20%, preferably about 100%, of the full-length antigen-induced interferon-γ and / or interleukin-12 in the model assays disclosed herein.

Portions and other variants of the M. tuberculosis antigen can be calculated by synthetic or recombinant means. Synthetic polypeptides of less than about 100 amino acids, generally less than about 50 amino acids, can be calculated using techniques known in the art. For example, such polypeptides can be synthesized using commercially available solid phase techniques such as Merrifield solid phase synthesis, in which amino acids are added sequentially to the growing amino acid chain. Merrifield, J. Am. Chem. Soc. 85: 2149-2146, 1963. Devices for automated synthesis of polypeptides are commercially available from a source, such as Perkin Elmer / Applied Biosystems Division, Foster City, CA, and can be manipulated according to the manufacturer's instructions. Variants of natural antigens can be prepared using standard mutation techniques, such as site specific mutagenesis derived from oligonucleotides. Truncated polypeptides can be prepared by removing fragments of the DNA sequence using standard techniques.

Recombinant polypeptides comprising portions and / or variants of natural antigens can be readily prepared from DNA sequences encoding polypeptides using a variety of techniques known in the art. For example, the supernatant obtained from a suitable host / vector system that secretes recombinant protein into the culture medium can first be concentrated using a commercial filter. After concentration, the concentrate can be applied to a suitable purification matrix such as an affinity matrix or an ion exchange resin. Finally, one or more reversed phase HPLC steps can be used to further purify the recombinant protein.

Any of a variety of expression vectors known in the art can be used to express the recombinant polypeptides of the invention. It can be expressed in any suitable host cell transformed or transfected with an expression vector comprising a DNA molecule encoding a recombinant polypeptide. Suitable host cells include prokaryotic cells, yeast and higher eukaryotic cells. The host cell used is preferably E. coli, yeast or mammalian cells such as COS or CHO. DNA sequences expressed in this manner can encode naturally occurring antigens, portions of naturally occurring antigens, or variants thereof.

In general, the polypeptides disclosed herein are prepared in substantially pure form, regardless of the method of preparation. The polypeptide is preferably at least about 80% pure, more preferably at least about 90% pure, and most preferably at least about 99% pure. In certain preferred embodiments, substantially pure polypeptides are incorporated into pharmaceutical compositions or vaccines for use in one or more methods disclosed herein, as described in detail below.

In an embodiment, the invention comprises (a) the DNA sequences of SEQ ID NOs: 1-12, 83, 102-108, 125, 127-137, 139 and 140; (b) an M. tuberculosis antigen (or a complementary sequence of this DNA sequence) or (c) at least one amino acid sequence encoded by a DNA sequence substantially homologous to the sequence of (a) or (b) Polypeptides comprising at least immunogenic portions of such antigens). In a related embodiment, the invention provides a minimal immunogenic portion of an M. tuberculosis antigen having an amino acid sequence selected from the group consisting of the sequences provided in SEQ ID NOs: 16-33, 109, 126, 138, 141, 142 and variants thereof It provides a polypeptide comprising a.

The M. tuberculosis antigens provided herein include variants that encode DNA sequences that are substantially homologous to one or more DNA sequences specifically described herein. As used herein, "substantial homology" refers to a DNA sequence that can hybridize under moderate stringent conditions. Suitable moderate stringent conditions include preliminary washing in 5 × SSC, 0.5% SDS, 1.0 mM EDTA (pH 8.0) solution; Hybridization at 50-65 ° C., 5 × SSC overnight, or 45 ° C., 0.5 × SSC for species homology; And two washes at 65 ° C. for 20 minutes with 2 × SSC, 0.5 × SSC and 0.2 × SSC containing 0.1% SDS, respectively. If such hybridized DNA sequences are nucleotides that encode immunogenic polypeptides that the hybridized DNA sequence encodes due to code degeneracy, this sequence also belongs to the present invention.

In a related aspect, the invention relates to fusion proteins comprising the first and second polypeptides of the invention, or alternatively to the polypeptides of the invention and known M. tuberculosis antigens such as Andersen and Hansen et al. Infect. Immun. 57: 2481-2488, 1989 (Genbank Accession No. M30046), or M. antigen. Vorbis (Accession No. U34848) and M. Provided is a fusion protein comprising ESAT-6 previously identified in tuberculosis (Sorensen et al., Infec. Immun. 63: 1710-1717, 1995). Variants of such fusion proteins are also provided. The fusion protein of the invention may comprise a linker peptide between the first polypeptide and the second polypeptide.

DNA sequences encoding the fusion proteins of the invention are constructed using known recombinant DNA techniques to associate separate DNA sequences encoding the first polypeptide and the second polypeptide into an appropriate expression vector. The 3 'end of the DNA sequence encoding the first polypeptide is ligated to the 5' end of the DNA sequence encoding the second polypeptide, irrespective of the presence of the peptide linker, so that mRNAs of the two DNA sequences can be ligated into the first polypeptide and the first polypeptide. There is a reading frame of the sequence in the translation into a single fusion protein that retains the biological activity of the two polypeptides.

Peptide linker sequences can be used to separate the first polypeptide and the second polypeptide at a distance sufficient to overlap each polypeptide with a secondary structure and a tertiary structure. Such peptide linker sequences are bottled into the fusion protein using standard techniques known in the art. Suitable peptide linker sequences can be selected based on the following factors. (1) the ability to form flexible elongated forms; (2) lack of ability to form secondary structures on the first polypeptide and the second polypeptide that can interact with functional epitopes; (3) Lack of hydrophobic or charged residues capable of reacting with polypeptide functional epitopes. Examples of preferred peptide linker sequences are Gly, Asn and Ser residues. Other approximate natural amino acids such as Thr and Ala can be used in the linker sequence. Amino acid sequences that can be usefully used as linkers are described in Maratea et al., Gene 40: 39-46, 1985; Murphy et al., Proc. Natl. Acad. Sci. USA 83: 8258-8262, 1986, US Pat. No. 4,935,233 and US Pat. No. 4,751,180. The linker sequence may be about 1 to about 50 amino acids in length. If the first polypeptide and the second polypeptide have a non-essential N terminal amino acid region that can be used to separate functional domains and prevent steric hindrance, no peptide sequence is required.

The ligated DNA sequence is operably linked to appropriate transcriptional or translational control components. The regulatory component responsible for DNA expression is located only 5 'to the DNA sequence encoding the first polypeptide. Similarly, the stop codon is only at 3 'to the DNA sequence encoding the second polypeptide.

In another aspect, the invention provides a method of inducing prophylactic immunity against tuberculosis in a patient using one or more of said polypeptides or fusion proteins (or DNA molecules encoding such polypeptides). "Patient" herein means any warm-blooded animal, preferably human. The patient may suffer from a disease or may be free of detectable diseases and / or infections. In other words, tuberculosis can be prevented or treated by inducing prophylactic immunity.

In this aspect, the polypeptide, fusion protein or DNA molecule is generally present in the pharmaceutical composition and / or vaccine. The pharmaceutical composition may comprise one or more polypeptides, and each polypeptide may comprise one or more such sequences (or variants thereof) and a physiologically acceptable carrier. The vaccine may comprise one or more of said polypeptides and nonspecific immune response enhancers such as adjuvants or liposomes with the polypeptide incorporated therein. Such pharmaceutical compositions and vaccines may comprise other M. tuberculosis antigens, either by incorporation into a combination polypeptide or by being present in a separate polypeptide.

Alternatively, the vaccine may comprise DNA encoding one or more of the polypeptides described above such that the polypeptides can be produced in situ. In such vaccines, the DNA may be present in any of a variety of delivery systems known in the art, including any nucleic acid expression system, bacterial expression system, and viral expression system. Suitable nucleic acid expression systems include DNA sequences necessary for expression in the patient (eg, appropriate promoter and termination signal). Bacterial delivery systems include administering bacteria (eg, Bacillus-Calmet-Guerin) that express an immunogenic portion of a polypeptide on the cell surface. In a preferred embodiment, the DNA can be introduced using a viral expression system (e.g. vaccinia or other pox virus, retrovirus or adenovirus) and the virus expression system can be used for non-pathogenic (incomplete), replication competent virus. It may include. Techniques for bottling DNA into such expression systems are known in the art. DNA is described, for example, as described in Ulmer et al. (Science 259: 1745-1749, 1993) and as reviewed in Cohen et al. (Science 259: 1691-1692, 1993). "Can be. Naked DNA uptake can be increased by coating DNA on biodegradable beads, which are effectively transported onto cells.

In a related aspect, the aforementioned DNA vaccines can be administered simultaneously or sequentially with the polypeptides of the invention or known M. tuberculosis antigens, such as the 38 kD antigen described above. For example, the DNA encoding the polypeptide of the invention may be administered in an "out of condition" or delivery system as described above, followed by administration of an antigen to enhance the prophylactic immune effect of the vaccine.

Dosages vary from subject to subject, as well as the route and frequency of administration, and can be used in the same manner as are currently used for immunization with BCG. In general, pharmaceutical compositions and vaccines may be administered by injection (eg, subcutaneously, intramuscularly, intravenously or subcutaneously), intranasally (eg, aspiration) or orally. Dosages can be administered from 1 to 3 times for 1 to 36 weeks. Three doses are administered at 3-4 month intervals, followed by periodic vaccination. Another protocol may be appropriate for individual patients. Appropriate doses are those amounts of polypeptides or DNA capable of raising the immune response in an immunized patient sufficient to protect the patient from M. tuberculosis infection for 1-2 years when administered as described above. Generally, the amount of polypeptide present in the dose (or co-generated by DNA in the dose) is from about 1 pg to about 100 mg, typically from about 10 pg to about 1 mg, preferably from about 100 pg to 1 kg of host. About 1 μg. Appropriate dose sizes may vary depending on the size of the patient, but are typically from about 0.1 ml to about 5 ml.

Any suitable carrier known to those skilled in the art can be used in the pharmaceutical compositions of the present invention, but the type of carrier depends on the mode of administration. For parenteral administration, such as subcutaneous infusion, the carrier preferably comprises water, saline, alcohol, lipids, waxes or buffers. For oral administration, any of the foregoing or solid carriers such as mannitol, lactose, starch, magnesium stearate, sodium saccharin, talc, cellulose, glucose, sucrose and magnesium carbonate can be used. Biodegradable microspheres (eg polylactic galactide) can be used as carriers for the pharmaceutical compositions of the present invention. Suitable biodegradable microspheres are disclosed in US Pat. Nos. 4,897,268 and 5,075,109.

Any of a variety of adjuvants can be used in the vaccines of the invention to nonspecifically enhance the immune response. Most adjuvants include substances designed to protect the antigen from rapid catabolism, such as sodium hydroxide or mineral oil, and nonspecific stimulants of the immune response, such as lipid A, Bortadella pertussis or Mycobacterium tubercules. Mycobacterium tuberculosis. Suitable adjuvants are commercially available, such as, for example, Freund's incomplete aid and Freund's complete adjuvant (Diffco Laboratories) and Merck Adjuvant 65 (Laway Merck and Company, NJ). Other suitable adjuvants include alum, biodegradable microspheres, monophosphate lipid A and quill A.

In another aspect, the invention provides a method of using the one or more polypeptides described above to identify tuberculosis using a skin test. As used herein, “skin test” is any assay that is performed directly in a patient by measuring delayed-type hypersensitivity (DTH) responses (eg, swelling, redness, dermatitis) following intradermal injection of one or more of the polypeptides described above. . Such injection may be performed using any device sufficient to contact the patient's skin cells with the polypeptide (s), such as a tuberculin syringe or a 1 ml syringe. The reaction is preferably measured at least 48 hours after injection, more preferably 48 to 72 hours after injection.

The DTH response is a cell mediated immune response, which is greater in patients who have previously been exposed to the test antigen (ie, the immunogenic portion of the polypeptide used or a variant thereof). The response can be measured visually using a ruler. In general, a response of at least about 0.5 cm in diameter, preferably at least about 1.0 cm, is a positive response indicating that it is a tuberculosis infection and may or may not be apparent as an active disease.

The polypeptide of the invention is preferably formulated for use in skin testing as a pharmaceutical composition comprising the polypeptide as described above and a physiologically acceptable carrier. Such compositions typically comprise one or more such polypeptides in an amount of about 1 μg to about 100 μg, preferably about 10 to 50 μg, in a volume of 0.1 ml. Carriers used in such pharmaceutical compositions are saline solutions with suitable preservatives such as phenol and / or Tween 80 ^™ .

In a preferred embodiment, the polypeptide used in the skin test is large enough to remain at the injection site for the duration of the reaction. In general, polypeptides of 9 or more amino acids in length are sufficient. In addition, the polypeptide is preferably degraded by macrophages within several hours of injection and provided to T cells. Such polypeptides may comprise repeats of said one or more sequences and / or other immunogenic or non-immunogenic sequences.

The following examples are provided for illustration only and are not intended to limit the invention.

Summary of the Invention

Briefly described, the present invention provides compositions and methods for the prevention and identification of tuberculosis. In a first aspect, the invention provides a polypeptide comprising an immunogenic portion of an M. tuberculosis antigen, or a variant of said antigen that has been altered only by conservative substitutions and / or modifications, wherein said antigen is SEQ ID NO: 1 , 11, 12, 83, 103-108, 125, 127, 129-137, 139 and 140, the complementary sequences of these sequences, and SEQ ID NOs: 1, 11, 12, 83, 103-108, 125, 127 And amino acid sequences encoded by DNA sequences selected from the group consisting of DNA sequences that hybridize to the sequences disclosed in 129-137, 139, and 140, or complementary sequences thereof, under ordinary stringent conditions. In a second aspect, the invention provides an immunogenic portion of an M. tuberculosis antigen having an amino acid sequence selected from the group consisting of the sequences provided in SEQ ID NOs: 16-33, 109, 126, 138, 141, 142 and variants thereof. It provides a polypeptide comprising.

In a related third aspect, there is provided a DNA sequence encoding a polypeptide as described above, an expression vector comprising these DNA sequences, and a host cell transformed or transfected with the expression vector.

In a fourth aspect, the invention provides a fusion protein comprising a first polypeptide and a second polypeptide of the invention, or alternatively a fusion protein comprising a polypeptide of the invention and a known M. tuberculosis antigen. will be.

In a fifth aspect, the present invention provides a pharmaceutical composition comprising at least one of said polypeptides, or a DNA molecule encoding such a polypeptide, and a physiologically acceptable carrier. In addition, the present invention provides a vaccine containing at least one polypeptide described above and a nonspecific immune response enhancer together with a vaccine containing at least one DNA sequence and a nonspecific immune response enhancer encoding such polypeptide.

In a sixth aspect, the present invention provides a method of inducing prophylactic immunity of a patient comprising administering to the patient an effective amount of at least one of said polypeptides.

In a seventh aspect of the invention, a method and a confirmation kit for detecting tuberculosis in a patient are provided. The method comprises contacting the at least one polypeptide with skin cells of the patient and detecting an immune response on the skin of the patient. The identification kit includes a device sufficient to contact the polypeptide with the skin cells of the patient with one or more of said polypeptides.

In an eighth aspect, the present invention provides a method for detecting tuberculosis of a patient, the method comprising the sequences set forth in SEQ ID NOs: 2-10, 102, 128, the complementary sequences of these sequences, and SEQ ID NOs: 2-10, 102, Contacting skin cells of a patient with at least one polypeptide encoded by a DNA sequence selected from the group consisting of DNA sequences hybridizing to the sequences disclosed in 128; And detecting an immune response in the skin of the patient. Identification kits for use in this method are also provided.

These and other aspects of the present invention will become apparent with reference to the following detailed description of the invention and the accompanying drawings. The entire text of each documented individually is incorporated herein by reference.

Example 1

Purification and Characterization of M. tuberculosis Polypeptides Using CD4 + T Cell Lines from Human PBMC

M. tuberculosis antigens of the present invention are substantially described in Sanderson et al. J. Exp. Med., 1995, 182: 1751-1757, cDNA libraries of M. tuberculosis strains H37Rv and Erdman were isolated by expression cloning, which contained IFN-γ and in immunoreactive T cell lines. It has been shown to induce PBMC proliferation.

Dendritic cells were infected with M. tuberculosis to create two CD4 + T cell lines called DC-4 and DC-5. Specifically, dendritic cells were prepared from adherent PBMCs obtained from a single donor and then infected with tuberculosis. Lymphocytes from the same donor were incubated with infected dendritic cells under limiting dilution conditions to produce CD4 + T cell lines DC-4 and DC-5. This cell line was observed to react with crude soluble protein from M. tuberculosis but not Tb38-1. Again, limiting dilution conditions were used to obtain a third CD4 + T cell line called DC-6, which was observed to react with both crude soluble protein and Tb38-1.

Genomic DNA was isolated from the M. tuberculosis strains H37Rv and Erdman, and this was used to construct an expression library in the vector pBSK (-) using lambda ZAP expression system (Stratazine, La Jolla, Calif.). Used. The library was transformed with E. coli, and the resulting collection of E. coli cultures were incubated with dendritic cells, and the resulting incubated dendritic cells were subjected to cell proliferation of the CD4 + T cell line DC-6. The nature of stimulating IFN- [gamma] production was investigated as described in Example 2 below. Positive collections were fractionated and retested until pure M. tuberculosis clones were obtained. Nineteen clones were isolated, nine of which were found to contain the previously identified M. tuberculosis antigens TbH-9 and Tb38-1, disclosed in US patent application 08 / 533,634. The sequenced cDNA sequences of the remaining 10 clones (hereinafter, Tb224, Tb636, Tb424, Tb436, Tb398, Tb508, Tb441, Tb475, Tb488 and Tb465) are described in SEQ ID NOs: 1-10, respectively. Corresponding amino acid sequences of Tb 224 and Tb636 are set forth in SEQ ID NOs: 13 and 14, respectively. The open reading frame of these two antigens was found to have some homology with TbH-9. In addition, Tb224 and Tb636 were found to be overlapping clones.

In addition, it has been found that Tb424, Tb436, Tb398, Tb508, Tb441, Tb475, Tb488 and Tb465 each contain two small open reading frames (called ORF-1 and ORF-2) or truncations thereof. The presence of micro variants of ORF-1 and ORF-2 in each clone was also confirmed. The deduced amino acid sequences corresponding to ORF-1 and ORF-2 of Tb424, Tb436, Tb398, Tb508, Tb441, Tb475, Tb488, and Tb465 are shown in SEQ ID NOs: 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26 and 27, 28 and 29, and 30 and 31 respectively. In addition, clones Tb424 and Tb436 were found to explicitly include a third open reading frame called ORF-U. The amino acid sequences inferred for the ORF-U of Tb424 and Tb436 are set forth in SEQ ID NOs: 32 and 33, respectively. Tb424 and Tb436 have been found to be overlapping clones or similar overlap / transpose copies. Similarly, Tb398, Tb508, and Tb465, like Tb475 and Tb488, were found to be overlapping clones or similar overlapping / transpose copies.

These sequences were compared to the known sequences of Genbank using the BLASTN system. No sequence homology with antigens Tb224 and Tb431 was found. Tb636 was found to be 100% identical to the cosmid identified in conventional M. tuberculosis. Similarly, Tb508, Tb488, Tb398, Tb424, Tb436, Tb441, Tb465 and Tb475 have also been found to be homologous to known M. tuberculosis cosmids. In addition, Tb488 was found to be 100% homologous to M. tuberculosis topoisomerase I.

17 overlapping peptides for the open reading frame ORF-1 (referred to as 1-1 to 1-17, SEQ ID NOs: 34-50) and 30 overlapping peptides for the open reading frame ORF-2 (hereafter 2-1 to 2) Called -30, SEQ ID NOs: 51-80) were synthesized according to the procedure described in Example 3 below.

The synthetic peptides, recombinant ORF-1 and ORF-2, induce IFN- [gamma] production and T cell proliferation in PBMCs of PPD positive donors were analyzed as described in Example 2 below. 1A and 1B and 2A and 2B illustrate T cell proliferation and stimulation of IFN-γ by recombinant ORF-2 and synthetic peptides 2-1 to 2-16 of two donors, called D7 and D160, respectively. Recombinant ORF-2 (hereinafter referred to as MTI) stimulated T cell proliferation and IFN-γ production in PBMCs of both donors. The amount of PBMC stimulation observed by each synthetic peptide was different for each donor, suggesting that each donor recognizes several epitopes on ORF2. The proteins encoded by ORF-1, ORF-2 and ORF-U were called MTS, MTI and MSF, respectively.

18 overlapping peptides (MSF-1 to MSF-18, SEQ ID NOs: 84 to 101, respectively) for the sequence of MSF were synthesized, and T cell proliferation in CD4 + T cell lines generated against M. tuberculosis culture filtrate, and Each property that stimulates IFN- [gamma] production was investigated as described below. Peptides called MSF-12 and MSF-13 (SEQ ID NOS: 95 and 96, respectively) were found to exhibit the highest reactivity. In addition, two overlapping peptides (SEQ ID NOS: 81 and 82) for the open reading frame of Tb224 were synthesized and found to induce T cell proliferation and IFN-γ production in PBMCs derived from PPD positive donors.

Two CD4 + T cell lines from different donors were prepared for M. tuberculosis infected dendritic cells according to the method described above. Using this cell line, the above-mentioned M. tuberculosis cDNA expression library was selected, and two clones called Tb867 and Tb391 were isolated. The cDNA sequence of the sequenced Tb867 was found to be identical to the previously isolated M. tuberculosis cosmid SCY22G10, and the candidate reactive open reading frame encoded the 750 amino acid M. tuberculosis protein kinase. Comparing the sequenced cDNA sequence of Tb391 with that of Genbank showed no significant homology with the known sequence.

As another study, a CD4 + T cell line was prepared for M. tuberculosis culture filtrate substantially as described above and used to select the M. tuberculosis Erdman cDNA expression library described above. Five reactive clones, Tb431, Tb472, Tb470, Tb838 and Tb962, were isolated. The sequenced cDNA sequences of sequenced Tb431, Tb472, Tb470 and Tb838 are set forth in SEQ ID NOs: 11, 12, 104 and 105, respectively, and the sequenced cDNA sequences of Tb962 are set forth in SEQ ID NOs: 106 and 107. The deduced amino acid sequence corresponding to Tb431 is shown in SEQ ID NO: 15.

Next, an experiment was performed to separate the full-length cDNA sequence (SEQ ID NO: 108) of Tb472. Overlapping peptides were synthesized and used to identify reactive open reading frames. The deduced amino acid sequence of the protein encoded by Tb472 (called MSL) is set forth in SEQ ID NO: 109. Comparing the sequences of Tb472 and MSL with those of Genbank as described above, it was found that there are no known sequences homologous. 15 overlapping peptides for MSL sequences (hereinafter referred to as MSL-1 to MSL-15, respectively; SEQ ID NOs: 110-124) were synthesized and T cells in CD4 + T cell lines generated against M. tuberculosis culture filtrates. Properties that stimulate proliferation and IFN- [gamma] production were investigated as described below. Peptides called MSL-10 (SEQ ID NO: 119) and MSL-11 (SEQ ID NO: 120) were found to exhibit the highest reactivity.

The cDNA sequence of the sequenced Tb838 was compared with the sequence in Genbank and found to be identical to the previously isolated M. tuberculosis cosmid SCY07H7. A comparison of the sequenced cDNA sequence of clone Tb962 and the sequence of Genbank revealed that it had some homology with two conventionally identified M. tuberculosis cosmids, one of which encodes some bactoferritin. . However, recombinant bactoferritin was found to be inactive with the T cell line used to isolate Tb962.

The clone Tb470 described above was used to recover a full-length open reading frame (SEQ ID NO: 125) homologous to TbH9 and found to encode a 40 kDa antigen called Mtb40. The amino acid sequence of the sequenced Mtb40 is set forth in SEQ ID NO: 126. Similarly, the full length cDNA sequence of Tb431 described in SEQ ID NO: 83 was then isolated and observed to include an open reading frame encoding Mtb40. In addition, Tb470 and Tb431 were found to contain potential open reading frames encoding U-ORF-like antigens.

M. tuberculosis Erdman cDNA expression libraries were selected from a plurality of CD4 + T cell lines generated against M. tuberculosis culture filtrate to isolate three clones, Tb366, Tb433 and Tb439. The cDNA sequences of the sequenced Tb366, Tb433 and Tb439 are set forth in SEQ ID NOs: 127, 128 and 129, respectively. Comparing this sequence with that of Jenbank, no sequence with significant homology to Tb366 was found. Tb433 was found to have some homology with the previously identified M. tuberculosis antigen MPT83. Tb439 was found to be 100% identical to the previously identified M. tuberculosis cosmid SCY02B10.

As described above, CD4 + T cell lines were prepared against M. tuberculosis PPD and used to select the M. tuberculosis Erdman cDNA expression library. One reactive clone (called Tb372) was isolated and sequenced cDNA sequences are set forth in SEQ ID NOs: 130 and 131. Comparing this sequence with that of Jenbank, no sequence with significant homology was found.

Subsequently, M. tuberculosis cDNA expression libraries were selected with CD4 + T cell lines generated for the dendritic cells infected with tuberculosis for 8 days as described above, and two clones, Tb390R5C6 and Tb390R2C11, were isolated. The cDNA sequence of the sequenced Tb390R5C6 is set forth in SEQ ID NO: 132 and the cDNA sequence of Tb390R2C11 is set forth in SEQ ID NOs: 133 and 134. Tb390R5C6 was found to be 100% identical to the previously identified M. tuberculosis cosmid.

The M. tuberculosis genomic DNA library prepared as follows was then selected using the method described above. Genomic DNA of the M. tuberculosis Erdman strain was randomly cut to an average size of 2 kb and blunt-terminated with Clinau polymerase, followed by addition of an EcoRI adapter. This insert was ligated to a screen phage vector (Novagen, Madison, WI) and packaged in vitro using phagemaker extract (Novagen). Phage libraries (referred to as Er λ screen libraries) were amplified and some were converted to plasmid expression libraries using E. coli strain BM25.8 (Novagen) as a self-subcloning mechanism. Plasmid DNA was purified from BM25.8 cultures containing pSCREEN recombinants and used to transform competent cells of the expression host strain BL21 (DE3) pLysS. Transformed cells were sized to have about 50 colonies per well in 96 well microtiter plates. Replica plates of 96 well plasmid library type were induced with IPTG to express recombinant proteins. After induction, the plates were centrifuged to precipitate E. coli and used directly for cloning T cell expression of CD4 + T cell lines made with the PPD positive donor (donor 160) described above. Collections containing E. coli expressing M. tuberculosis T cell antigen were isolated into individual colonies and then reanalyzed in a similar manner to identify positive groups.

T cell lines from donor 160 were selected with one 96 well plate of the Erdλ screen library to obtain a total of nine positive groups. Conventional experiments selecting pBSK libraries described above with T cells from donor 160 indicate that most or all of the positive clones will be TbH-9, Tb38-1 or MTI (disclosed in US Patent Application 08 / 533,634) or a variant thereof. Implied. However, Southern analysis revealed that only three wells hybridize with mixed probes of TbH-9, Tb38-1 and MTI. Two of the remaining six positive wells were found to be identical. The 5 'cDNA sequences sequenced for two isolated clones (Y1-26C1 and Y1-86C11) are shown in SEQ ID NOs: 135 and 136, respectively. The full length cDNA sequence of the isolated clone, called hTcc # 1, is shown in SEQ ID NO: 137 and the corresponding inferred amino acid sequence is shown in SEQ ID NO: 138. Comparing the sequence of hTcc # 1 and the sequence of Genbank as described above, it was found that there is some homology with the previously identified M. tuberculosis MTCY07H7B.06.

Example 2

Induction of T cell proliferation and interferon-γ production by M. tuberculosis antigen

Recombinant M. tuberculosis antigen was measured as follows whether the activity to induce T cell proliferation and interferon-γ production.

Proteins are described in Skeiky et al. J. Exp. Med., 1995, 181: 1527-1537, induced by IPTG and purified by gel elution. Purified polypeptides were then screened for inducing T cell proliferation in PBMC preparations. PBMCs, known to be positive for the PPD skin test and known to proliferate T cells in response to PPD, were cultured in a medium containing RPMI 1640 supplemented with 10% collected human serum and 50 μg / ml of gentamicin. Purified polypeptide was added in duplicates at a concentration of 0.5-10 μg / ml. Six days after incubation at a volume of 200 μl in a 96 well round bottom plate, 50 μl of medium was isolated from each well and the amount of IFN-γ was measured as described below. This plate was then pulsed with tritiated thymidine 1 μCi / well for 18 hours, harvested, and the tritium uptake was measured with a gas scintillation counter. Fractions with at least three-fold proliferation in both replicates were considered positive than proliferation observed in cells cultured in media alone.

IFN- [gamma] was measured using an enzyme bound immunosorbent assay (ELISA). ELISA plates were coated for 4 hours at room temperature with mouse monoclonal antibodies directed to human IFN- [gamma] contained in PBS (PharMingen, San Diego, Calif.). The wells were then blocked with PBS containing 5% (W / V) skim milk powder for 1 hour at room temperature. The plates were washed six times with PBS / 0.2% TWEEN-20, diluted 1: 2 with the culture medium and injected into ELISA plates and incubated overnight at room temperature. This plate was washed again and polyclonal rabbit antihuman IFN- [gamma] serum diluted 1: 3000 fold in PBS / 10% normal goat serum was added to each well. The plates were then incubated for 2 hours at room temperature and washed, and then horseradish peroxidase bound antirabbit IgG (Sigma Chemical Company, St. Louis, MO) was placed in PBS / 5% skim milk powder. Added in 1: 2000 dilution. After incubation for 2 hours at room temperature, the plates were washed and TMB substrate added. The reaction was stopped after 20 minutes with 1N sulfuric acid. The reference wavelength was 570 nm and the optical density was measured at 450 nm. It was considered positive if the OD of the fractions obtained from both replicates was at least 2 times and the standard deviation was 3 compared to the mean OD of cells cultured only in the medium alone.

Example 3

Purification and Characterization of M. tuberculosis polypeptides using CD4 + T cell line prepared from mouse M. tuberculosis model

C57BL / 6 mice were infected with M. tuberculosis to cause progressive disease for about 2-3 weeks. The progression of this disease was stopped by forming a strong prophylactic T cell mediated immune response. This infection model was used to create a T cell line capable of recognizing the prophylactic M. tuberculosis antigen.

Specifically, splenocytes were isolated from C57BL / 6 mice infected with M. tuberculosis for 28 days to induce specific anti-M. Tuberculosis T cell lines as described above. The above-mentioned M. tuberculosis Erdλ screen library was selected using normal antigen expression (spleen) cells from C57BL / 6 mice with the resulting CD4 + T cell line. One of the reactive library collections found to be highly stimulating against T cells was selected and the corresponding active clones (called Y288C10) were isolated.

Sequencing clone Y288C10 was found to contain two potential genes side by side. The cDNA sequences (called mTCC # 1 and mTCC # 2) of the two genes sequenced are shown in SEQ ID NOs: 139 and 140, respectively, and the corresponding deduced amino acid sequences are shown in SEQ ID NOs: 141 and 142, respectively. This sequence was compared with that of Genbank and found to be identical to the previously known unknown sequence in M. tuberculosis cosmid MTY21C12. The deduced amino acid sequences of mTCC # 1 and mTCC # 2 were found to be slightly homologous to members of the previously identified TbH9 protein family as described above.

Example 4

Synthesis of Synthetic Polypeptides

Polypeptides were synthesized with a Millipore 9050 peptide synthesizer using FMOC chemistry with HPTU (O-benzotriazole-N, N, N ', N'-tetramethyluronium hexafluorophosphate) activation. The Gly-Cys-Gly sequence was attached to the amino terminus of the peptide to provide a method for conjugation or labeling of the peptide. Cleavage of the peptide from the solid support can be carried out using the following cleavage mixture, trifluoroacetic acid: ethanedithiol: thioanisol: water: phenol (40: 1: 2: 2: 3). After cleavage for 2 hours, the peptide can be precipitated in cold methyl-t-butyl-ether. The peptide pellets were then dissolved in water containing 0.1% trifluoroacetic acid (TFA), lyophilized and purified by C18 reversed phase HPLC. Peptides were eluted using acetonitrile (containing 0.1% TFA) gradient 0-60% in water (containing 0.1% TFA). After lyophilization of the pure fractions, the peptides were characterized by electrospray mass spectrometry and amino acid analysis.

The present invention has been described in detail through specific embodiments of the present invention, and various modifications may be made without departing from the spirit and scope of the present invention.

Sequence list

(1) General Information:

(i) Applicants: Alderson, Mark; Dillon, Davin C. Skekey, Yash A. W .; Campos-Neto, Antonio

(ii) Name of the invention: Compound for immunotherapy and identification of tuberculosis and use thereof

(iii) Number of sequences: 144

(iv) Letter address:

(A) Recipient: Seed and Berry

(B) Street: 5th Avenue 701, Cumbia Center 6300

(C) city: Seattle

(D) State: Washington

(E) Country: United States

(F) ZIP code: 98104

(v) computer readable form:

(A) Medium type: floppy disk

(B) Computer: IBM PC compatible model

(C) operating system: PC-DOS / MS-DOS

(D) Software: PatentIn Release # 1.0, Version # 1.30

(vi) Current application data:

(A) Application number:

(B) Application date: May 5, 1998

(C) Classification symbol:

(viii) Patent Attorney / Agent Information:

(A) Name: Maki, David J.

(B) registration number: 31,392

(C) Reference / Control Number: 210121.440C1

(ix) Communication Information:

(A) Telephone: 206-622-4900

(B) Fax: 206-682-6031

(2) Information about SEQ ID NO: 1

(i) Sequence Properties:

(A) Length: 1886 base pairs

(B) Sequence type: nucleic acid

(C) Number of chains: 2 strands

(D) topology: linear

(ii) Type of molecule: DNA (genome)

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO 1:

[SEQ ID NO: 1]

(2) Information about SEQ ID NO:

(i) Sequence Properties:

(A) Length: 2305 base pairs

(B) Sequence type: nucleic acid

(C) Number of chains: 2 strands

(D) topology: linear

(ii) Type of molecule: DNA (genome)

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO: 2:

[SEQ ID NO 2]

(2) Information about SEQ ID NO:

(i) Sequence Properties:

(A) Length: 1742 base pairs

(B) Sequence type: nucleic acid

(C) Number of chains: 2 strands

(D) topology: linear

(ii) Type of molecule: DNA (genome)

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO: 3:

[SEQ ID NO 3]

(2) Information about SEQ ID NO:

(i) Sequence Properties:

(A) Length: 2836 base pairs

(B) Sequence type: nucleic acid

(C) Number of chains: 2 strands

(D) topology: linear

(ii) Type of molecule: DNA (genome)

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO 4:

[SEQ ID NO 4]

(2) Information about SEQ ID NO:

(i) Sequence Properties:

(A) Length: 900 base pairs

(B) Sequence type: nucleic acid

(C) Number of chains: 2 strands

(D) topology: linear

(ii) Type of molecule: DNA (genome)

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO: 5:

[SEQ ID NO: 5]

(2) Information about SEQ ID NO:

(i) Sequence Properties:

(A) Length: 1905 base pairs

(B) Sequence type: nucleic acid

(C) Number of chains: 2 strands

(D) topology: linear

(ii) Type of molecule: DNA (genome)

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO 6:

[SEQ ID NO: 6]

(2) Information about SEQ ID NO:

(i) Sequence Properties:

(A) Length: 2921 base pairs

(B) Sequence type: nucleic acid

(C) Number of chains: 2 strands

(D) topology: linear

(ii) Type of molecule: DNA (genome)

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO 7:

[SEQ ID NO: 7a]

[SEQ ID NO: 7b]

(2) Information about SEQ ID NO: 8

(i) Sequence Properties:

(A) Length: 1704 base pairs

(B) Sequence type: nucleic acid

(C) Number of chains: 2 strands

(D) topology: linear

(ii) Type of molecule: DNA (genome)

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO: 8:

[SEQ ID NO: 8a]

[SEQ ID NO: 8b]

(2) Information about SEQ ID NO:

(i) Sequence Properties:

(A) Length: 2286 base pairs

(B) Sequence type: nucleic acid

(C) Number of chains: 2 strands

(D) topology: linear

(ii) Type of molecule: DNA (genome)

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO: 9:

[SEQ ID NO: 9a]

[SEQ ID NO: 9b]

(2) Information about SEQ ID NO: 10:

(i) Sequence Properties:

(A) Length: 1136 base pairs

(B) Sequence type: nucleic acid

(C) Number of chains: 2 strands

(D) topology: linear

(ii) Type of molecule: DNA (genome)

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO: 10:

[SEQ ID NO: 10]

(2) Information about SEQ ID NO:

(i) Sequence Properties:

(A) Length: 967 base pairs

(B) Sequence type: nucleic acid

(C) Number of chains: 2 strands

(D) topology: linear

(ii) Type of molecule: DNA (genome)

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO: 11:

[SEQ ID NO: 11]

(2) Information about SEQ ID NO: 12:

(i) Sequence Properties:

(A) Length: 585 base pairs

(B) Sequence type: nucleic acid

(C) Number of chains: 2 strands

(D) topology: linear

(ii) Type of molecule: DNA (genome)

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO: 12:

[SEQ ID NO: 12]

(2) Information about SEQ ID NO:

(i) Sequence Properties:

(A) Length: 144 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO: 13:

[SEQ ID NO: 13]

(2) Information about SEQ ID NO: 14:

(i) Sequence Properties:

(A) Length: 352 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO: 14:

[SEQ ID NO: 14]

(2) Information about SEQ ID NO: 15:

(i) Sequence Properties:

(A) Length: 141 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO: 15:

[SEQ ID NO: 15]

(2) Information about SEQ ID NO: 16:

(i) Sequence Properties:

(A) Length: 58 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO 16:

[SEQ ID NO: 16]

(2) Information about SEQ ID NO: 17:

(i) Sequence Properties:

(A) Length: 67 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO: 17:

[SEQ ID NO: 17]

(2) Information about SEQ ID NO: 18:

(i) Sequence Properties:

(A) Length: 58 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO: 18:

[SEQ ID NO: 18]

(2) Information about SEQ ID NO: 19:

(i) Sequence Properties:

(A) Length: 94 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO: 19:

[SEQ ID NO: 19]

(2) Information about SEQ ID NO: 20:

(i) Sequence Properties:

(A) Length: 30 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO: 20:

[SEQ ID NO: 20]

(2) Information about SEQ ID NO: 21:

(i) Sequence Properties:

(A) Length: 94 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO: 21:

[SEQ ID NO: 21]

(2) Information about SEQ ID NO: 22:

(i) Sequence Properties:

(A) Length: 69 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO: 22:

[SEQ ID NO: 22]

(2) Information about SEQ ID NO: 23:

(i) Sequence Properties:

(A) Length: 94 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO: 23:

[SEQ ID NO: 23]

(2) Information about SEQ ID NO: 24:

(i) Sequence Properties:

(A) Length: 52 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO: 24:

[SEQ ID NO: 24]

(2) Information about SEQ ID NO: 25:

(i) Sequence Properties:

(A) Length: 94 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO: 25:

[SEQ ID NO: 25]

(2) Information about SEQ ID NO: 26:

(i) Sequence Properties:

(A) Length: 98 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO: 26:

[SEQ ID NO: 26]

(2) Information about SEQ ID NO: 27:

(i) Sequence Properties:

(A) Length: 94 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO: 27:

[SEQ ID NO: 27]

(2) Information about SEQ ID NO: 28:

(i) Sequence Properties:

(A) Length: 81 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO: 28:

[SEQ ID NO: 28]

(2) Information about SEQ ID NO: 29:

(i) Sequence Properties:

(A) Length: 94 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO: 29:

[SEQ ID NO: 29]

(2) Information about SEQ ID NO: 30:

(i) Sequence Properties:

(A) Length: 11 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO: 30:

[SEQ ID NO: 30]

(2) Information about SEQ ID NO: 31:

(i) Sequence Properties:

(A) Length: 94 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO: 31:

[SEQ ID NO: 31]

(2) Information about SEQ ID NO: 32:

(i) Sequence Properties:

(A) Length: 99 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO: 32:

[SEQ ID NO: 32]

(2) Information about SEQ ID NO: 33:

(i) Sequence Properties:

(A) Length: 99 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO: 33:

[SEQ ID NO: 33]

(2) Information about SEQ ID NO: 34:

(i) Sequence Properties:

(A) Length: 15 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO: 34:

[SEQ ID NO: 34]

(2) Information about SEQ ID NO: 35:

(i) Sequence Properties:

(A) Length: 15 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO: 35:

[SEQ ID NO: 35]

(2) Information about SEQ ID NO: 36:

(i) Sequence Properties:

(A) Length: 15 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO: 36:

[SEQ ID NO: 36]

(2) Information about SEQ ID NO: 37:

(i) Sequence Properties:

(A) Length: 15 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO: 37:

[SEQ ID NO: 37]

(2) Information about SEQ ID NO: 38:

(i) Sequence Properties:

(A) Length: 15 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO: 38:

[SEQ ID NO: 38]

(2) Information about SEQ ID NO: 39:

(i) Sequence Properties:

(A) Length: 15 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO: 39:

[SEQ ID NO: 39]

(2) Information about SEQ ID NO: 40:

(i) Sequence Properties:

(A) Length: 15 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO: 40:

[SEQ ID NO: 40]

(2) Information about SEQ ID NO: 41:

(i) Sequence Properties:

(A) Length: 16 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO: 41:

[SEQ ID NO: 41]

(2) Information about SEQ ID NO: 42:

(i) Sequence Properties:

(A) Length: 15 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO: 42:

[SEQ ID NO: 42]

(2) Information about SEQ ID NO: 43:

(i) Sequence Properties:

(A) Length: 15 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO: 43:

[SEQ ID NO: 43]

(2) Information about SEQ ID NO: 44:

(i) Sequence Properties:

(A) Length: 15 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO: 44:

[SEQ ID NO: 44]

(2) Information about SEQ ID NO: 45:

(i) Sequence Properties:

(A) Length: 15 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO: 45:

[SEQ ID NO: 45]

(2) Information about SEQ ID NO: 46:

(i) Sequence Properties:

(A) Length: 15 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO: 46:

[SEQ ID NO: 46]

(2) Information about SEQ ID NO: 47:

(i) Sequence Properties:

(A) Length: 15 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO: 47:

[SEQ ID NO: 47]

(2) Information about SEQ ID NO: 48:

(i) Sequence Properties:

(A) Length: 15 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO: 48:

[SEQ ID NO 48]

(2) Information about SEQ ID NO: 49:

(i) Sequence Properties:

(A) Length: 16 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO: 49:

[SEQ ID NO: 49]

(2) Information about SEQ ID NO: 50:

(i) Sequence Properties:

(A) Length: 17 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO: 50:

[SEQ ID NO 50]

(2) Information about SEQ ID NO: 51:

(i) Sequence Properties:

(A) Length: 15 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO: 51:

[SEQ ID NO: 51]

(2) Information about SEQ ID NO: 52:

(i) Sequence Properties:

(A) Length: 15 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO: 52:

[SEQ ID NO: 52]

(2) Information about SEQ ID NO: 53:

(i) Sequence Properties:

(A) Length: 15 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO: 53:

[SEQ ID NO: 53]

(2) Information about SEQ ID NO: 54:

(i) Sequence Properties:

(A) Length: 15 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO: 54:

[SEQ ID NO: 54]

(2) Information about SEQ ID NO: 55:

(i) Sequence Properties:

(A) Length: 15 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO: 55:

[SEQ ID NO: 55]

(2) Information about SEQ ID NO: 56:

(i) Sequence Properties:

(A) Length: 15 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO: 56:

[SEQ ID NO: 56]

(2) Information about SEQ ID NO: 57:

(i) Sequence Properties:

(A) Length: 15 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO: 57:

[SEQ ID NO: 57]

(2) Information about SEQ ID NO: 58:

(i) Sequence Properties:

(A) Length: 16 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO: 58:

[SEQ ID NO: 58]

(2) Information about SEQ ID NO: 59:

(i) Sequence Properties:

(A) Length: 15 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO: 59:

[SEQ ID NO: 59]

(2) Information about SEQ ID NO: 60:

(i) Sequence Properties:

(A) Length: 15 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO: 60:

[SEQ ID NO: 60]

(2) Information about SEQ ID NO: 61:

(i) Sequence Properties:

(A) Length: 18 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO: 61:

[SEQ ID NO: 61]

(2) Information about SEQ ID NO: 62:

(i) Sequence Properties:

(A) Length: 15 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO: 62:

[SEQ ID NO: 62]

(2) Information about SEQ ID NO: 63:

(i) Sequence Properties:

(A) Length: 15 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO: 63:

[SEQ ID NO: 63]

(2) Information about SEQ ID NO: 64:

(i) Sequence Properties:

(A) Length: 15 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO: 64:

[SEQ ID NO: 64]

(2) Information about SEQ ID NO: 65:

(i) Sequence Properties:

(A) Length: 15 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO: 65:

[SEQ ID NO: 65]

(2) Information about SEQ ID NO: 66:

(i) Sequence Properties:

(A) Length: 16 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO: 66:

[SEQ ID NO: 66]

(2) Information about SEQ ID NO: 67:

(i) Sequence Properties:

(A) Length: 15 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO: 67:

[SEQ ID NO: 67]

(2) Information about SEQ ID NO: 68:

(i) Sequence Properties:

(A) Length: 15 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO: 68:

[SEQ ID NO: 68]

(2) Information about SEQ ID NO: 69:

(i) Sequence Properties:

(A) Length: 15 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO: 69:

[SEQ ID NO: 69]

(2) Information about SEQ ID NO: 70:

(i) Sequence Properties:

(A) Length: 15 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO: 70:

[SEQ ID NO: 70]

(2) Information about SEQ ID NO: 71:

(i) Sequence Properties:

(A) Length: 15 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO: 71:

[SEQ ID NO: 71]

(2) Information about SEQ ID NO: 72:

(i) Sequence Properties:

(A) Length: 15 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO: 72:

[SEQ ID NO: 72]

(2) Information about SEQ ID NO: 73:

(i) Sequence Properties:

(A) Length: 15 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO: 73:

[SEQ ID NO: 73]

(2) Information about SEQ ID NO: 74:

(i) Sequence Properties:

(A) Length: 15 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO: 74:

[SEQ ID NO: 74]

(2) Information about SEQ ID NO: 75:

(i) Sequence Properties:

(A) Length: 15 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO: 75:

[SEQ ID NO: 75]

(2) Information about SEQ ID NO: 76:

(i) Sequence Properties:

(A) Length: 15 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO: 76:

[SEQ ID NO: 76]

(2) Information about SEQ ID NO: 77:

(i) Sequence Properties:

(A) Length: 16 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO: 77:

[SEQ ID NO: 77]

(2) Information about SEQ ID NO: 78:

(i) Sequence Properties:

(A) Length: 15 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO: 78:

[SEQ ID NO: 78]

(2) Information about SEQ ID NO: 79:

(i) Sequence Properties:

(A) Length: 15 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO: 79:

[SEQ ID NO: 79]

(2) Information about SEQ ID NO: 80:

(i) Sequence Properties:

(A) Length: 15 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO: 80:

[SEQ ID NO: 80]

(2) Information about SEQ ID NO: 81:

(i) Sequence Properties:

(A) Length: 25 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO: 81:

[SEQ ID NO: 81]

(2) Information about SEQ ID NO: 82:

(i) Sequence Properties:

(A) Length: 25 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(vi) origin:

(A) Biological name: Mycobacterium tuberculosis

(xi) SEQ ID NO: 82:

[SEQ ID NO: 82]

(2) Information about SEQ ID NO: 83:

(i) Sequence Properties:

(A) Length: 967 base pairs

(B) Sequence type: nucleic acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: cDNA

(xi) SEQ ID NO: 83:

[SEQ ID NO: 83]

(2) Information about SEQ ID NO: 84:

(i) Sequence Properties:

(A) Length: 15 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(xi) SEQ ID NO: 84:

[SEQ ID NO: 84]

(2) Information about SEQ ID NO: 85:

(i) Sequence Properties:

(A) Length: 15 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(xi) SEQ ID NO: 85:

[SEQ ID NO: 85]

(2) Information about SEQ ID NO: 86:

(i) Sequence Properties:

(A) Length: 15 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(xi) SEQ ID NO: 86:

[SEQ ID NO: 86]

(2) Information about SEQ ID NO: 87:

(i) Sequence Properties:

(A) Length: 15 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(xi) SEQ ID NO: 87:

[SEQ ID NO: 87]

(2) Information about SEQ ID NO: 88:

(i) Sequence Properties:

(A) Length: 15 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(xi) SEQ ID NO: 88:

[SEQ ID NO: 88]

(2) Information about SEQ ID NO: 89:

(i) Sequence Properties:

(A) Length: 15 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(xi) SEQ ID NO: 89:

[SEQ ID NO: 89]

(2) Information about SEQ ID NO: 90:

(i) Sequence Properties:

(A) Length: 15 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(xi) SEQ ID NO: 90:

[SEQ ID NO: 90]

(2) Information about SEQ ID NO: 91:

(i) Sequence Properties:

(A) Length: 15 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(xi) SEQ ID NO: 91:

[SEQ ID NO: 91]

(2) Information about SEQ ID NO: 92:

(i) Sequence Properties:

(A) Length: 15 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(xi) SEQ ID NO: 92:

[SEQ ID NO: 92]

(2) Information about SEQ ID NO: 93:

(i) Sequence Properties:

(A) Length: 15 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(xi) SEQ ID NO: 93:

[SEQ ID NO: 93]

(2) Information about SEQ ID NO: 94:

(i) Sequence Properties:

(A) Length: 15 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(xi) SEQ ID NO: 94:

[SEQ ID NO: 94]

(2) Information about SEQ ID NO: 95:

(i) Sequence Properties:

(A) Length: 15 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(xi) SEQ ID NO: 95:

[SEQ ID NO: 95]

(2) Information about SEQ ID NO: 96:

(i) Sequence Properties:

(A) Length: 16 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(xi) SEQ ID NO: 96:

[SEQ ID NO: 96]

(2) Information about SEQ ID NO: 97:

(i) Sequence Properties:

(A) Length: 15 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(xi) SEQ ID NO: 97:

[SEQ ID NO: 97]

(2) Information about SEQ ID NO: 98:

(i) Sequence Properties:

(A) Length: 15 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(xi) SEQ ID NO: 98:

[SEQ ID NO: 98]

(2) Information about SEQ ID NO: 99:

(i) Sequence Properties:

(A) Length: 15 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(xi) SEQ ID NO: 99:

[SEQ ID NO: 99]

(2) Information about SEQ ID NO: 100:

(i) Sequence Properties:

(A) Length: 15 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(xi) SEQ ID NO: 100:

[SEQ ID NO: 100]

(2) Information about SEQ ID NO: 101:

(i) Sequence Properties:

(A) Length: 14 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(xi) SEQ ID NO: 101:

[SEQ ID NO: 101]

(2) Information about SEQ ID NO: 102:

(i) Sequence Properties:

(A) Length: 1784 base pairs

(B) Sequence type: nucleic acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: cDNA

(xi) SEQ ID NO: 102:

[SEQ ID NO: 102a]

[SEQ ID NO: 102b]

(2) Information about SEQ ID NO: 103:

(i) Sequence Properties:

(A) Length: 766 base pairs

(B) Sequence type: nucleic acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: cDNA

(xi) SEQ ID NO: 103:

[SEQ ID NO: 103]

(2) Information about SEQ ID NO: 104:

(i) Sequence Properties:

(A) Length: 1231 base pairs

(B) Sequence type: nucleic acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: cDNA

(xi) SEQ ID NO: 104:

[SEQ ID NO: 104]

(2) Information about SEQ ID NO: 105:

(i) Sequence Properties:

(A) Length: 2041 base pairs

(B) Sequence type: nucleic acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: cDNA

(xi) SEQ ID NO: 105:

[SEQ ID NO: 105]

(2) Information about SEQ ID NO: 106:

(i) Sequence Properties:

(A) Length: 1202 base pairs

(B) Sequence type: nucleic acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: cDNA

(xi) SEQ ID NO: 106:

[SEQ ID NO: 106]

(2) Information about SEQ ID NO: 107:

(i) Sequence Properties:

(A) Length: 496 base pairs

(B) Sequence type: nucleic acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: cDNA

(xi) SEQ ID NO: 107:

[SEQ ID NO: 107]

(2) Information about SEQ ID NO: 108:

(i) Sequence Properties:

(A) Length: 849 base pairs

(B) Sequence type: nucleic acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: cDNA

(xi) SEQ ID NO: 108:

[SEQ ID NO: 108]

(2) Information about SEQ ID NO: 109:

(i) Sequence Properties:

(A) Length: 97 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: protein

(xi) SEQ ID NO: 109:

[SEQ ID NO: 109]

(2) Information about SEQ ID NO: 110:

(i) Sequence Properties:

(A) Length: 15 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(xi) SEQ ID NO: 110:

[SEQ ID NO: 110]

(2) Information about SEQ ID NO: 111:

(i) Sequence Properties:

(A) Length: 15 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(xi) SEQ ID NO: 111:

[SEQ ID NO: 111]

(2) Information about SEQ ID NO: 112:

(i) Sequence Properties:

(A) Length: 15 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(xi) SEQ ID NO: 112:

[SEQ ID NO: 112]

(2) Information about SEQ ID NO: 113:

(i) Sequence Properties:

(A) Length: 15 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(xi) SEQ ID NO: 113:

[SEQ ID NO: 113]

(2) Information about SEQ ID NO: 114:

(i) Sequence Properties:

(A) Length: 15 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(xi) SEQ ID NO: 114:

[SEQ ID NO: 114]

(2) Information about SEQ ID NO: 115:

(i) Sequence Properties:

(A) Length: 15 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(xi) SEQ ID NO: 115:

[SEQ ID NO: 115]

(2) Information about SEQ ID NO: 116:

(i) Sequence Properties:

(A) Length: 15 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(xi) SEQ ID NO: 116:

[SEQ ID NO: 116]

(2) Information about SEQ ID NO: 117:

(i) Sequence Properties:

(A) Length: 15 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(xi) SEQ ID NO: 117:

[SEQ ID NO: 117]

(2) Information about SEQ ID NO: 118:

(i) Sequence Properties:

(A) Length: 15 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(xi) SEQ ID NO: 118:

[SEQ ID NO: 118]

(2) Information about SEQ ID NO: 119:

(i) Sequence Properties:

(A) Length: 15 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(xi) SEQ ID NO: 119:

[SEQ ID NO: 119]

(2) Information about SEQ ID NO: 120:

(i) Sequence Properties:

(A) Length: 15 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(xi) SEQ ID NO: 120:

[SEQ ID NO: 120]

(2) Information about SEQ ID NO: 121:

(i) Sequence Properties:

(A) Length: 15 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(xi) SEQ ID NO: 121:

[SEQ ID NO: 121]

(2) Information about SEQ ID NO: 122:

(i) Sequence Properties:

(A) Length: 15 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(xi) SEQ ID NO: 122:

[SEQ ID NO: 122]

(2) Information about SEQ ID NO: 123:

(i) Sequence Properties:

(A) Length: 15 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(xi) SEQ ID NO: 123:

[SEQ ID NO: 123]

(2) Information about SEQ ID NO: 124:

(i) Sequence Properties:

(A) Length: 18 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: peptide

(xi) SEQ ID NO: 124:

[SEQ ID NO: 124]

(2) Information about SEQ ID NO: 125:

(i) Sequence Properties:

(A) Length: 1752 base pairs

(B) Sequence type: nucleic acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: cDNA

(xi) SEQ ID NO: 125:

[SEQ ID NO: 125]

(2) Information about SEQ ID NO: 126:

(i) Sequence Properties:

(A) Length: 400 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: protein

(xi) SEQ ID NO: 126:

[SEQ ID NO: 126]

(2) Information about SEQ ID NO: 127:

(i) Sequence Properties:

(A) Length: 474 base pairs

(B) Sequence type: nucleic acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: cDNA

(xi) SEQ ID NO: 127:

[SEQ ID NO: 127]

(2) Information about SEQ ID NO: 128:

(i) Sequence Properties:

(A) Length: 1431 base pairs

(B) Sequence type: nucleic acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: cDNA

(xi) SEQ ID NO: 128:

[SEQ ID NO: 128]

(2) Information about SEQ ID NO: 129:

(i) Sequence Properties:

(A) Length: 279 base pairs

(B) Sequence type: nucleic acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: cDNA

(xi) SEQ ID NO: 129:

[SEQ ID NO: 129]

(2) Information about SEQ ID NO: 130:

(i) Sequence Properties:

(A) Length: 1470 base pairs

(B) Sequence type: nucleic acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: cDNA

(xi) SEQ ID NO: 130:

[SEQ ID NO: 130]

(2) Information about SEQ ID NO: 131:

(i) Sequence Properties:

(A) Length: 1059 base pairs

(B) Sequence type: nucleic acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: cDNA

(xi) SEQ ID NO: 131:

[SEQ ID NO: 131]

(2) Information about SEQ ID NO: 132:

(i) Sequence Properties:

(A) Length: 153 base pairs

(B) Sequence type: nucleic acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: cDNA

(xi) SEQ ID NO: 132:

[SEQ ID NO: 132]

(2) Information about SEQ ID NO: 133:

(i) Sequence Properties:

(A) Length: 387 base pairs

(B) Sequence type: nucleic acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: cDNA

(xi) SEQ ID NO: 133:

[SEQ ID NO: 133]

(2) Information about SEQ ID NO: 134:

(i) Sequence Properties:

(A) Length: 389 base pairs

(B) Sequence type: nucleic acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: cDNA

(xi) SEQ ID NO: 134:

[SEQ ID NO: 134]

(2) Information about SEQ ID NO: 135:

(i) Sequence Properties:

(A) Length: 480 base pairs

(B) Sequence type: nucleic acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: cDNA

(xi) SEQ ID NO: 135:

[SEQ ID NO: 135]

(2) Information about SEQ ID NO: 136:

(i) Sequence Properties:

(A) Length: 587 base pairs

(B) Sequence type: nucleic acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: cDNA

(xi) SEQ ID NO: 136:

[SEQ ID NO: 136]

(2) Information about SEQ ID NO: 137:

(i) Sequence Properties:

(A) Length: 1200 base pairs

(B) Sequence type: nucleic acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: cDNA

(xi) SEQ ID NO: 137:

[SEQ ID NO: 137]

(2) Information about SEQ ID NO: 138:

(i) Sequence Properties:

(A) Length: 392 base pairs

(B) Sequence type: nucleic acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: protein

(xi) SEQ ID NO: 138:

[SEQ ID NO: 138]

(2) Information about SEQ ID NO: 139:

(i) Sequence Properties:

(A) Length: 439 base pairs

(B) Sequence type: nucleic acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: cDNA

(xi) SEQ ID NO: 139:

[SEQ ID NO: 139]

(2) Information about SEQ ID NO: 140:

(i) Sequence Properties:

(A) Length: 1441 base pairs

(B) Sequence type: nucleic acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: cDNA

(xi) SEQ ID NO: 140:

[SEQ ID NO: 140a]

[SEQ ID NO: 140b]

(2) Information about SEQ ID NO: 141:

(i) Sequence Properties:

(A) Length: 99 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: protein

(xi) SEQ ID NO: 141:

[SEQ ID NO: 141]

(2) Information about SEQ ID NO: 142:

(i) Sequence Properties:

(A) Length: 423 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: protein

(xi) SEQ ID NO: 142:

[SEQ ID NO: 142a]

[SEQ ID NO: 142b]

(2) Information about SEQ ID NO: 143:

(i) Sequence Properties:

(A) Length: 97 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: protein

(xi) SEQ ID NO: 143:

[SEQ ID NO: 143]

(2) Information about SEQ ID NO: 144:

(i) Sequence Properties:

(A) Length: 99 amino acids

(B) Sequence type: amino acid

(C) Number of chains: 1 chain

(D) topology: linear

(ii) Type of molecule: protein

(xi) SEQ ID NO: 144:

[SEQ ID NO: 144]

Claims (29)

The sequences set forth in SEQ ID NOs: 1, 11, 12, 83, 103-108, 125, 127, 129-137, 139 and 140, the complementary sequences of these sequences, and SEQ ID NOs: 1, 11, 12, 83, 103-108, M. tubercule, comprising an amino acid sequence encoded by a DNA sequence selected from the group consisting of DNA sequences that hybridize to the sequences disclosed in 125, 127, 129-137, 139, and 140, or their complementary sequences, under normal stringent conditions A polypeptide comprising an immunogenic portion of a M. tuberculosis antigen, or a variant of said antigen changed only by conservative substitutions and / or modifications. An immunogenic portion of an M. tuberculosis antigen, or only conservative substitutions, comprising an amino acid sequence selected from the group consisting of the sequence set forth in SEQ ID NOs: 16-33, 109, 126, 138, 141, 142 and variants thereof And / or a polypeptide comprising a variant of said antigen altered by a modification. A DNA molecule comprising a nucleotide sequence encoding the polypeptide of claim 1. An expression vector comprising the DNA molecule of claim 3. A host cell transformed with the expression vector according to claim 4. 6. The host cell of claim 5, wherein said host cell is selected from the group consisting of E. coli, yeast and mammalian cells. A pharmaceutical composition comprising at least one polypeptide of claim 1 or a physiologically acceptable carrier. A pharmaceutical composition comprising at least one DNA molecule of claim 3 and a physiologically acceptable carrier. A pharmaceutical composition comprising at least one DNA molecule having a sequence set forth in SEQ ID NOs: 2-10, 102, 128 and a physiologically acceptable carrier. A vaccine comprising at least one polypeptide of claim 1 or a nonspecific immune response enhancer. 1 type encoded by a DNA sequence selected from the group consisting of a sequence disclosed in SEQ ID NOs: 2-10, 102, 128, a complementary sequence of this sequence, and a DNA sequence hybridizing to the sequence disclosed in SEQ ID NOs: 2-10, 102, 128 Or more polypeptides; And A vaccine comprising a nonspecific immune response enhancer. The vaccine of claim 10 or 11, wherein the nonspecific immune response enhancer is an adjuvant. A vaccine comprising at least one DNA molecule of claim 3 and a nonspecific immune response enhancer. A vaccine comprising at least one DNA molecule having a sequence set forth in SEQ ID NOs: 2-10, 102, 128 and a nonspecific immune response enhancer. The vaccine of claim 13 or 14, wherein the nonspecific immune response enhancer is an adjuvant. A method of inducing prophylactic immunity in a patient, comprising administering to the patient the pharmaceutical composition of claim 7. A method of inducing prophylactic immunity in a patient comprising administering to the patient the vaccine of claim 10. A fusion protein comprising two or more kinds of polypeptides according to claim 1. A fusion protein comprising the polypeptide of claim 1 or a known M. tuberculosis antigen. A pharmaceutical composition comprising the fusion protein of claim 18 or 19 and a physiologically acceptable carrier. A vaccine comprising the fusion protein of claim 18 or 19 and a nonspecific immune response enhancer. The vaccine of claim 21, wherein the nonspecific immune response enhancer is an adjuvant. A method of inducing prophylactic immunity in a patient, comprising administering the pharmaceutical composition of claim 20 to the patient. A method of inducing prophylactic immunity in a patient comprising administering to the patient the vaccine of claim 21. (a) contacting a skin cell of a patient with at least one polypeptide according to claim 1; And (b) detecting the immune response on the skin of the patient, and detecting tuberculosis in the patient therefrom. (a) a DNA sequence selected from the group consisting of a sequence disclosed in SEQ ID NOs: 2-10, 102, 128, a complementary sequence of this sequence, and a DNA sequence hybridizing to the sequence disclosed in SEQ ID NOs: 2-10, 102, 128 Contacting the patient's skin cells with one or more polypeptides; And (b) detecting the immune response on the skin of the patient, and detecting tuberculosis in the patient therefrom. 27. The method of claim 25 or 26, wherein the immune response is cirrhosis. (a) a polypeptide according to claim 1 or 2; And (b) a confirmation kit comprising a device sufficient to contact the skin cells of the patient with the polypeptide. (a) a DNA sequence selected from the group consisting of a sequence disclosed in SEQ ID NOs: 2-10, 102, 128, a complementary sequence of this sequence, and a DNA sequence hybridizing to the sequence disclosed in SEQ ID NOs: 2-10, 102, 128 Polypeptide; And (b) a confirmation kit comprising a device sufficient to contact the skin cells of the patient with the polypeptide.